Over one-half of the global population is expected to be obese by 2030 [1]. Obesity is a physically and psychologically debilitating cycle that is positively correlated with depression symptoms and incidental depression, implying that further weight gain will only worsen symptoms [2]. These sorts of mental health implications further the notion that obesity is not only a physically debilitating condition, but a psychological one as well. While causes of obesity and excessive weight gain may vary, the symptoms and biological markers are rather consistent. Grossly excess white adipose tissue composition is the primary factor relating to the function and potential remedial effects of BAT reprogramming [3]. Furthermore, hypertension, inflammation, and insulin resistance should be noted as telling symptoms associated with obesity.

Brown adipocytes are known to host an increased number of mitochondria, especially relative to white adipocytes [4]. The function of the mitochondria in this instance varies slightly from their “quintessential'' function, as these organelles operate to produce heat via non-shivering thermogenesis. Attenuation of ATP synthesis is due to a principal gene, uncoupling protein 1 (UCP-1), one whose products dissipate the coordination of the electron transport chain [5]. By rerouting protons through a channel alternative to ATP synthase, energy produced via oxidative phosphorylation is dissipated as heat as opposed to generating ATP. Furthermore, BAT activity is stimulated by cold temperatures and noradrenaline [6].

Our lab has previously demonstrated that adenoviral co-expression of HB-EGF and ADAM-12S in mammalian cells results in transdifferentiation of white adipose tissue (WAT) into BAT-like cells. These findings may serve as a potential combatant for obesity and type II diabetes. 

Primary research thus far has indicated potentially differentially regulated genes when comparing WT HEK293 cells to HB-EGF/ADAM12-S co-infected HEK293 cells. 

What genes might be differentially regulated?

What mechanisms are these genes associated with?

How do their differential expression levels, respectively, coalesce about the nature of brown adipocytes?

An HB-EGF and ADAM12-S adenoviral co-expression vector was designed and implemented in HEK-293 co-transfection alongside mock-infected cells; conferring cellular transdifferentiation was done by Oil Red-O staining, nuclear and mitochondrial fluorescence, and analysis of the cells’ proliferative state. Total RNA isolation was performed on the HEK-293 cells after two weeks of infection. Genomic DNA was then eliminated, followed by reverse transcription to synthesize cDNA which was then added to SYBR Green fluorescent dye and aliquoted into a 96-well qPCR Array. Subsequently, real-time polymerase chain reaction (RT-PCR) was performed to determine differential gene expression using Qiagen RT2 Profiler Obesity array. 

Evidence of the gene products should be pursued to verify downregulation of these factors. A western blot will be used about TNFα, employing polyclonal antibodies to observe the protein products. 

Nearsighted research might determine whether the reprogrammed cells are able to substantiate metabolism with a preference for lipids, as opposed to simple sugars.

Obesity therapies should be considered in the form of BAT reprogramming, relative to pertinent processes: metabolic preference, insulin resistance, Type-2 Diabetes, and induced-autoimmune diseases

The research conducted in this lab has fostered my drive for higher education and problem solving. The skills I have acquired in this lab have surely prepared me for graduate-level engagements. Barring quantitative analysis, the qualitative component of my involvement has allowed me to hypothesize about our results and design potential experiments, which has only piqued my interest and creativity further. Dr. Harding abets mentoring relationships with his undergraduates, which facilitates positive feedback and progressive thinking to further our findings and substantiate our claims about this cutting edge research. Being an upperclassman on this project has provided me the opportunity to assist and guide some of the upcoming researchers that are involved in this project. The collaborative nature of our lab implies teamwork in a constant effort to accomplish common goals. Our research team has been introduced and become proficient in a multitude of scientific practices, while versing ourselves in scientific etiquette and professionalism. Furthermore, presenting at the ASCB conference in San Diego (2024) provided me a chance to witness the world of research first-hand. Although daunting, this experience provided me a degree of insight into the professional realm of research and the requisite dedication to academia that may just change the world. 

 [1] Finkelstein E, et al. (2012). AJPM. 42(6): 563-570. [2] Xin Guo Y, et al. (2023). BES. 36(6): 481-489. [3] Shi L, et al. (2022). Nat Metab 4: 1573–1590. [4] Cannon B, et al. (2004). Physiol Rev. 84: 277–359. [5] Porter R (2006). 1757(5–6): 446-448. [6] Cinti S (2006). 16(8): 569-574. [7] Hotamisligil GS (1999). 245: 621-625. [8] Marchand L, et al. (2002). 94(6): 454–460. [9] Dong L, et al. (2020). 19(4): 1513-1521